Preflight

what's our goal

Ian: "The chemicals we'll be using amount to waste water (E. coli), cheese whey (tryptone), Vegemite (yeast extract & NaCl), tofu firming salts (MgCl2 and MgSO4), jelly (agar) and antibiotics and the concentrations are fairly low, so no special dangers - i.e. don't eat it. Cleanup can be city sewer, since all of these are things that are already found in waste water."

what are the risks today to us

trivial; just doing a dry run w/ the water baths

what are the risks today to the community

water

what waste are we gonna generate

hot water; combusted propane gas

Protocol

Thaw competent cells (Lyocomp E. coli) on ice.

During this time pre-chill sterile centrifuge tubes on ice.

Gently tap competent cell containers to mix

Aliquot bacterial suspension in pre-chilled tubes (100 µl per tube).

To the aliquots add 1 µl of plasmid (RFP PUC19).

Gently tap centrifuge tubes to mix

Incubate 30 min on ice.

During this time pre-heat water bath to 42°C (don’t use heating block).